Syed M. Ali

Abstract 1592: Anti-cancer evaluation of various solvent extracts of blue honeysuckle berry (Lonicera caerulea L.) against prostate cancer cells

Prostate cancer (PCa) is the second most common cancer among men in the United Sates. It is estimated 1 in 6 men will be diagnosed with PCa and approximately 180,890 new cases are predicted in 2016 alone. Standard treatments for PCa include surgery, radiation, chemotherapy and hormonal therapy or a combination of these treatments. However, factors like patient health, drug resistance, specificity, and toxicity can result in poor disease prognosis. In order to overcome these limitations and improve patient prognosis, there is an urgent need to identify new anti-cancer agents with minimal side effects. Current studies are being focused on natural products and their components for alternative therapeutics. Blue Honeysuckle (Lonicera caerule L.) a berry native to northeast Asia, is known to be rich in Vitamin C and polyphenols such as anthocyanins, flavonoids, and phenolic acids. Polyphenols are found to have several therapeutic effects such as anti-inflammatory, antioxidant and antimicrobial properties. Our present study used sequential solvent extracts of Blue Honeysuckle (BHS) berry using Hexane, Ethyl Acetate, Methanol, and Water respectively. These fractions were used to assess various therapeutic effects of BHS on DU 145, PC-3, C4-2 and LNCaP PCa cell lines. The goal was to identify the most potent BHS fraction that is effective against PCa using pre-clinical studies. MTT assays were used to identify the anti-proliferative effects of various BHS fractions. The above indicated PCa cells were treated with different doses (10-150 µg/mL) of BHS factions over various time periods (24, 48, 72 hr). Our data revealed that Hexane extract (HE) exhibited the highest inhibition of cell viability in a time and dose-dependent manner. HE fraction showed to have an IC50 of 89.6 μg/mL for DU 145, 117.4 μg/mL for PC-3, 163.3 μg/mL for C4-2 and 140.84 for LNCaP cells. Moreover, BHS HE fraction showed a decrease in migration capacity and colony formation ability of PCa cells in vitro. Cellular senescence assay was performed to assess β-Galactosidase activity in PCa cells treated with BHS HE extract. All the PCa cell lines treated with 100 µg/mL of BHS HE showed increased senescence. Western blotting was performed to identify the potential anti-cancer mechanism of BHS against PCa using DU 145 and PC-3 cell lines. Results from this study indicated that DU 145 and PC-3 cell lines after 24 hr treatment with 100 μg/mL of various BHS fractions especially BHS HE showed an increase in apoptosis (Caspase 3,8,9 and PARP-1) and autophagy (LC3 A/B) markers. Furthermore, cell cycle analysis of DU 145 and PC-3 cells treated with BHS HE extract showed increased apoptotic cells. Further analysis of apoptotic and autophagy pathways modulated by BHS treatments are required to illustrate its potential underlying anti-cancer mechanisms in PCa. In conclusion, results from our study warrant further evaluation of BHS berry for potential PCa management. Citation Format: Syed M. Ali, Alex Ourth, Chun-Tao Che, Mian-Ying Wang, Gnanasekar Munirathinam. Anti-cancer evaluation of various solvent extracts of blue honeysuckle berry (Lonicera caerulea L.) against prostate cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1592. doi:10.1158/1538-7445.AM2017-1592

Abstract 3905: Synthesis and structure activity relationship of substituted N,6-diphenyl-5,6-dihydrobenzo[h]quinazolin-2-amine as inhibitors of fibroblast growth factor receptors (FGFR)

Utilization of hydrophobic motifs present in auto-inhibited protein kinases has resulted in the identification of a series of 5,6-dihydrobenzo [h]quinazolin-2-amines with activity as fibroblast growth factor receptor (FGFR) tyrosine kinase inhibitors. Herein we describe the combination of a proprietary in silico design process, a new screening paradigm using an array of biochemical and biophysical technologies in conjunction with an established parallel chemistry process for the identification and optimization of a series of novel FGFR inhibitors. These potent FGFR inhibitors exhibit a preference for the inactive form of the kinase, are non-ATP competitive, and exhibit robust cellular pharmacodynamic inhibition as well as in vitro anti-proliferative effects in cells dependent on FGFR and significant anti-tumor activity in appropriate xenograft models in vivo. The design strategy, synthesis, structure activity relationships and in vitro and in vivo biology of selected inhibitors will be presented. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3905. doi:1538-7445.AM2012-3905

Abstract 2645: Customized PEG linkers improve the pharmaceutical properties of cytotoxic small molecules

Introduction: PEGylation is an established delivery technology for proteins with benefits such as decreased immunogenicity and prolonged circulating half-life. The application of PEGylation to small molecules may improve the poor pharmaceutical properties including poor solubility, suboptimal pharmacokinetic (PK) profiles, and unwanted toxicities. Here we utilized Customized PEG Linkers to enhance the therapeutic index of several cytotoxic agents including doxorubicin, ara-C, gemcitabine, and SN38. Experimental procedures: drug molecules were either reacted with proper linker moieties first before PEGylation, or conjugated directly with PEG linkers. For those compounds thathave more than two reactive functional groups, the unwanted reactive sites were first protected before conjugation with PEG linkers. The PEG conjugates were incubated in PBS and plasma to study their stability, followed by in vivo PK studies in mice. Additionally, in vitro anti-proliferation assays with different human cancer cell lines were conducted to evaluate the PEG conjugates with different half-lives. Furthermore, the PEG conjugates were evaluated in vivo in multiple human cancer models in mice for their anticancer activities. Summary of data: a series of novel PEG conjugates were synthesized with different Customized PEG Linkers. In general, while these PEG conjugates were stable in PBS buffer, they demonstrated a broad range of half-lives in rat and human plasma, varying from minutes to days. PEG conjugates with longer half-lives have shown prolonged circulation time and increased AUC compared to native drugs in the PK studies in mice. In addition, PEGylation also greatly increased the water solubility for those insoluble molecules, for instance by about 1000-fold for SN38. In the cellular based studies, PEG conjugates showed different degrees of anticancer activities against a panel of human cancer cells. Remarkably, enhanced anticancer activities compared to the non-pegylated agent have been observed for many of the conjugates in a variety of cancer models, including both solid tumors and hematological malignancies. Conclusions: Customized PEG Linkers have greatly improved the solubility of certain small molecule drugs and enabled the systemic administration of these drugs. It allowed us to generate a variety of PEG-drug conjugates with broad half-life ranges in plasma which are clinically relevant. The combined effect of increased solubility, optimized PK profile, and passive accumulation of macromolecular PEG conjugates in tumors due to the enhanced permeation and retention effect may all contributed to the greatly enhanced anticancer efficacy of these PEG conjugates in animal models. In summary, customized PEG Linkers represent a promising technology to improve current cytotoxic agents. One agent, PEG-SN38 is undergoing Phase II evaluation in cancer patients with metastatic colorectal and breast cancer. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2645.